Objective To investigate the therapeutic effect of transplantation of limbal epithelial cells and decellularized porcine cornea construct in rabbits with corneal alkali burn.Methods Allogeneic rabbit limbal epithelial cells were cultured and identified.Porcine cornea scaffold was fabricated by decellularization.The LECs-DPC construct was cultured and transplanted to the alkali-burned corneal surface of rabbits(4month follow-up).Results Immunocytochemistry and RT-PCR analyses revealed the presence of ABCG2 positive limbal stem cells.A general view showed the corneas were clear and vessel-free postoperatively.HE and Masson Trichrome staining results indicated the epithelial stratification,fewer cellular infiltration,DPC' integration into host corneal tissues,and more regular collagen configuration.Transmission electron microscopy exhibited formation of desmosomes and microvilli.Conclusion Allogeneic limbal epithelial cells transplantation using DPC as a carrier can restore corneal clarity after alkali burn in a rabbit model.

Objective To observe the effect of MAC of the isoflurane at the general combined with epidural anesthesia with lidocaine and dicaine.Methods 78 cases of the cholecystectomy patients randomly double-blindly divided Ⅳ groups. General combined with epidural anesthesia were performed at the Ⅰ、Ⅱ、 Ⅲ groups. And general anesthesia were performed at the contronal group. The MAC of the isoflurane are checked with up-and-down method with electronic stimulation at left on C5 leveral. Results The MAC of the three observed groups of the general combined with epidural anaesthesia were 0.67?0.097%、0.68?0.084%、0.61?0.103 % differently. There is no siginificant difference.But the MAC of the general contronl group is 1.15?0.088%.There are siginificant difference with the three observed groups,The MAC of the isoflurane can be reduced with general combined with epidural anesthesia. Coinclousions The MAC the isoflurane can be reduced with general combined with epidural anaesthsia significantly compared with singal general anesthesia. And there are no significant different in the three observed groups.It is the pulsing action with lidocaine combined with dicaine at epidural anesthesia. The mechanism of anaesthesia with lidocaine or dicaine supposed to be the same.

BACKGROUND: Many scholars have paid attentions on neural stem cells (NSCs), which are regarded as the cell source to repair the injured nervous tissues. And it is becoming a key point that NSCs will be obtained in an effective way.OBJECTIVE: To observe the characteristics of isolation, proliferation and differentiation of in vitro NSCs from subventricular zone (SVZ) and hippocampus of neonatal rats.DESIGN: Single-sample trial.SETTING: Department of Neurosurgery in the Third Affiliated Hospital of Sun Yat-sen University.MATERIALS: Ten SD rats aged 3 days, of either gender, were offered from the Experimental Animal Center of Sun Yat-sen University. Nestin antibody (rabbit-anti-rat), neurofilament (NF-200) antibody (rabbit-anti-rat), and glial fibrillary acidic protein (GFAP) antibody (mice-anti-rat) were all purchased from Sigma Company.METHODS: The experiment was carried out in the Laboratory of Histology and Embryology, Basic Medical College of Sun Yat-sen University from September to December in 2006. According to the ethical requirement, the serum-free DMEM/F12 medium containing basic fibroblast growth factor and epidermal growth factor was utilized to isolate and incubate NSCs from SVZ and hippocampus of neonatal rats. Then the proliferation and differentiation of NSCs in vitro were observed.MAIN OUTCOME MEASURES: Fluorescence immunocytochemistry was applied to detect the antigen of nestin expressed from NSCs, NF expressed from neuron cells and GFAP expressed from astrocytes. The differentiation of NSCs was identified.RESULTS: The cells isolated from SVZ and hippocampus of neonatal rats possessed the ability of proliferation and self-renewal. Through inverted microscope, nestin positive cells could be found in the cell clone sphere at passage 20 of cells. Cells induced to differentiate could express antigen of NF and GFAP by the attached cell clone sphere.CONCLUSION: The NSCs successfully isolated from SVZ and hippocampi of neonatal rats possess the ability of self-renewal, proliferation, and differentiation into neuron-like cells and astrocytes.

Objective To explore the most important risk factors in metabolic syndrome(MS) components.Methods Ninety-four individuals were classified into MS and non-MS group according to the diagnostic criteria for MS proposed by Chinese Diabetes Society(CDS) revised in 2006 or International Diabetes Federal(IDF) in 2005.Age,waist circumference(WC),body mass index(BMI),fasting plasma glucose,lipid profile,blood pressure and blood cell counts in two groups were compared.Partial least squares discriminant analysis(PLSDA) was carried out to determine the most important components of MS.Results Patients with MS diagnosed by CDS or IDF criteria have significantly older age,higher BMI,WC,blood pressure,fasting plasma glucose,triglycerides,insulin levels,insulin resistance index,high sensitivity CRP and fibrinogen levels compared with non-MS group.PLSDA analysis shows WC,BMI,blood pressure and aging are most important components of MS.Conclusion Obesity,hypertension and aging are three most important components of MS with obesity is the utmost among them.

Objective To investigate the diagnostic value of the early slope value of enhancement in differentiating benign from malignant musculoskeletal masses.Methods Dynamic contrast-enhanced gradient-recalled acquisition in the steady state imaging(GRASS) was performed to evaluate 36 musculoskeletal masses(18 malignant,18 benign).The difference of the early slope value of enhancement was analysed and compared between benign lesions and malignant ones.Using ROC curve plotting obtained the best decisive threshold.Spearman rank correlation analysis was used to determine the relation between slope and microvessel density of tumor tissues.Results The average early slope value of enhancement was (4.33?2.70)% per second (range 0.97%～10.50% per second)for malignant lesions,and was (1.46?0.97)% per second(range 0.37%～3.86% per second)for benign ones.There was an apparent statistical significance between benign and malignant tumors(?

Objective To investigate the correlation of dynamic contrast-enhanced (DCE) MRI features with microvessel density (MVD) and vascular endothelial growth factor (VEGF) in non-small cell lung cancer(NSCLC).Methods Conventional MR imaging and dynamic contrast-enhanced scan in thirty-three patients with NSCLC confirmed by pathologyn were performed. MVD and VEGF were stained with immuno-histochemical technique in all cases. Some parameters of DCE MRI, including maximum slope(Smax) and time to peak(TTP) were put more analysis. The relationship between the results of DCE MRI (Smax and TTP) and that of immuno-histochemistry (MVD and VEGF) was analysed.Results The Smax of adeno carcinoma was higher than that of squamous cell carcinoma,but TTP was lower. The difference was obvious difference(t=3.22,P

Eleven type 1 diabetic patients who received fixed regime of insulin Glargine were included in the study.The levels of fasting serum insulin were measured for each subject at 6:00 in three consecutive mornings.The variability of mean fasting serum insulin in each subject was 3.3%-41.5% (mean 15.4%).The variability did not correlate with the dose of Glargine statistically.

Purpose To study the reversal effect on multidrug resistance (MDR) by TNF-α gene combined with verapamil (VRP) or tamoxifen (TAM). Methods By using recombinant retrovirus vector, TNF-α gene was transfected into multidrug-resistant human breast cancer cell line MCF7/ADR. The TNF-α secreting cell clone MCF7/ADR-TNF was obtained by G418 screening. The integrating and secreting of TNF-α were analyzed by PCR and ELISA. MTT assay and formula"I = d/D1 + d/D2" were used to evaluate the reversal effect of multidmg resistance with TNF-α gene combined with verapamil or tamoxifen. ResultsThe level of TNF-α secreted by MCF7/ADR-TNF was 1 737 pg/ml (106cells/48 h). Compared with control,the resistance to ADR of MCF7/ADR-TNF was reversed by 1.6 times. The reversal effect produced by combination of TNF-α gene and VRP was antagonistic. The combination of TNF-α gene and TAM produced synergic effect (interaction index I = 0.64). ConclusionsTNF-α gene combined with TAM has synergic effect on reversing MDR.

Objective To evaluate the activation of the visual cortex in patients with primary openangle glaucoma (POAG) and to explore whether the neuronal activity corresponds with retinal nerve fiber layer (RNFL) and cup-to-disc (C/D) values.Methods Twenty-five patients and 25 gender-and agematched healthy volunteers were studied.Blood oxygenation level-dependent functional magnetic resonance imaging (BOLD-fMRI) and three-dimensional brain volume imaging (3 D BRAVO) sequences were obtained using 3 T MR imaging system.A full-screen black-white shift checkerboard was used for visual stimulus during the fMRI experiment and was performed on each eye of all subjects using a visual-acoustical system.All acquired data were postprocessed and analyzed by statistical parametric mapping (SPM).After analysis,individual activated mapping,intra-group mean activated mapping,and inter-group variant mapping were observed.The voxel number,intensity,and Montreal Neurological Institute (MNI) coordinate of the activated clusters were recorded.The Xjviewer software was utilized to obtain activated voxel numbers in occipital lobe.A Pearson correlated test was performed to test the correlation between the number of activated voxels and RNFL,C/D and Hodapp-Anderson-Parrish (HAP) clinical stage.Results Intra-group mean activated mappings of both patients and volunteers showed obvious activation in bilateral occipital lobes.As compared with healthy volunteers,the POAG patients exhibited statistically significantly decreased activation in bilateral occipital lobes,left hippocampus,and left cerebellum,along with lower mean RNFL [(71.56 ±21.54) i m versus (111.88 ± 9.96) μm] and higher C/D values (0.71 ± 0.18 versus 0.36 ± 0.08 ; t value was respectively-10.901 and 11.643,P ＜ 0.05).The number of activated voxels in the occipital lobes of POAG patients did not correlate with RNFL,C/D and HAP clinical stage of the corresponding eye (r value was respectively 0.157,-0.113 and-0.242,P ＞ 0.05).Conclusions fMRI demonstrated differences in visual cortex activation in POAG patients relative to healthy volunteers,suggesting it might be a promising complementary method for diagnosing glaucoma.However,fMRI findings did not correlate with POAG extent,as measured by RNFL and C/D values.Ophthalmological examination remains to play an important role in the evaluation of open-angle glaucoma.

Objective To study the pathological alterations,such as oxidative stress,cell proliferation and insulin resistance,especially autophagy,in Alzheimer' s disease (AD) complicated with type 2 diabetes (AD + T2DM).Methods The mouse models of T2DM,AD and AD + T2DM were used in the study,and totally 80 mice were divided into four groups:control group,T2DM group,AD group and AD + T2DM group.Morris water maze was applied to test the ability of learning and memory among the above mentioned groups.In the meantime,insulin resistance index,the expression of insulin receptor substrate 2,oxidative stress,cell proliferation and autophagy were observed with chemical analysis,immunofluorescent labeling,transmission electron microscopy and Western blotting.Results On day 4,the difference of time to find Morris water maze in control group,T2DM group,AD group and AD + T2DM group ((26.08 ±4.93) s,(38.46 ± 4.07) s,(47.32 ± 5.86) s,(53.01 ± 6.12) s,F =2.454,P =0.025) was statistically significant,and the time in AD + T2DM group was longer than that in AD group (t =-3.624,P =0.033).Compared with control group,insulin resistance occurred in T2DM group,AD group and AD + T2DM group (4.35 ± 0.48,16.12 ± 3.57,7.03 ± 3.11,18.78 ± 5.06,F =5.602,P =0.009),and the reduction of insulin receptor substrate 2 expression,the oxidative stress reaction,neural proliferative suppression and autophagy (F =418.344,222.514,436.250,113.934,23.772,35.469,all P ＜ 0.05) were induced in T2DM group,AD group and AD + T2DM group,which were more serious in AD + T2DM group than in AD group (t =-2.796,21.723,-8.041,9.037,-4.403,-32.011,-26.593,all P ＜0.05).Conclusion AD + T2DM mice suffered more serious cognitive impairment than AD and T2DM mice.The oxidative stress levels of AD + T2DM mice were upregulated,and thus led to the inhibition of cell proliferation,eventually leading to promotion of autophagy.